Glycosynthases: Mutant glycosidases for oligosaccharide synthesis

Journal of the American Chemical Society

Volumn 120, Issue 22, 1998, Pages 5583-5584

  Mackenzie, Lloyd F ^a   Wang, Qingping ^a   Warren, R Antony J ^a   Withers, Stephen G ^a  

^a UNIVERSITY OF BRITISH COLUMBIA   (Canada)

Author keywords

[No Author keywords available]

Indexed keywords

GLYCOSIDASE; OLIGOSACCHARIDE; SYNTHETASE;

CANADA; CARBOHYDRATE SYNTHESIS; CATALYSIS; GLYCOSYLATION; LETTER; RHIZOBIUM; STEREOCHEMISTRY;

EID: 0032503519     PISSN: 00027863     EISSN: None     Source Type: Journal    
DOI: 10.1021/ja980833d     Document Type: Article

References (33)

1. Kobata, A. Acc. Chem. Res. 1993, 26, 319.
2. Varki, A. Glycobiology 1993, 3, 97.
3. Wong, C.-H.; Halcomb, R. L.; Ichikawa, Y.; Kajimoto, T. Angew. Chem., Int. Ed. Engl. 1995, 34, 521.
4. Sears, P.; Wong, C.-H. Proc. Natl. Acad. Sci. U.S.A. 1996, 93, 12086.
5. Zopf, D.; Roth, S. The Lancet 1996, 347, 1017.
6. Simon, P. M. Drug Discovery Today 1996, 1, 522.
7. Danishefsky, S. J.; Bilodeau, M. Angew. Chem., Int. Ed. 1996 35, 1380.
8. Boons, G.-J. Tetrahedron 1996, 52, 1095.
9. Douglas, S. P.; Whitfield, D. M.; Krepinsky, J. J. J Am. Chem. Soc. 1991, 113, 3, 5095.
10. Roberge, J. Y.; Beebe, X.; Danishefsky, S. J. Science 1995, 269, 202.
11. Liang, R.; Yan, L.; Loebach, J.; Ge, M.; Uozumi, Y.; Sekanina, K.; Horan, N.; Gildersleeve, J.; Thompson, C.; Smith, A.; Biswas, K.; Still, W. C.; Kahne, D. Science 1996, 274, 1520.
12. Nilsson, J. G. I. TIBTECH 1988, 6, 256. Thiem,
13. J. FEMS Microbiol. Rev. 1995, 16, 193.
14. Complex, but in many cases effective, recycling schemes can be used to regenerate NDPsugars in some cases (Ichikawa, Y.; Lin, Y.-C.; Dumas, D. P.; Shen, G.-J.; Garcia-Junceda, E.; Williams, M. A.; Bayer, R.; Ketcham, C.; Walker, L. E.; Paulson, J. C.; Wong, C.-H. J. Am. Chem. Soc. 1992,114, 9283).
15. Retaining glycosidases hydrolyze glycosides with net retention of anomeric configuration. Synthesis can also be carried out under equilibrium conditions, using high concentrations of the free sugars.
16. McCarter, J. D.; Withers, S. G. Curr. Opin. Struct. Biol. 1994, 4, 885-892.
17. Sinnott, M. L. Chem. Rev. 1990, 90, 1171.
18. m values they are used by the enzyme in preference to the accumulating products.
19. Equilibria have been displaced using a coupled enzyme to selectively convert product formed by adsorption of product onto charcoal and by selective crystallization of product or use of organic cosolvents.
20. Wakarchuk, W. W.; Campbell, R. L.; Sung, W. L.; Davoodi, J.; Yaguchi, M. Protein Sci. 1994, 3, 467.
21. Yuan, J.; Martinez-Bilbao, M.; Huber, R. E. Biochem. J. 1994, 299, 527.
22. Withers, S. G.; Rupitz, K.; Trimbur, D.; Warren, R. A. J. Biochemistry 1992, 31, 9979.
23. Wang, Q.; Graham, R. W.; Trimbur, D.; Warren, R. A. J.; Withers, S. G. J. Am. Chem. Soc. 1994, 116, 11594.
24. 12
25. 12
26. a of the acid/base catalyst 2.5 units, to a value below the optimum pH (McIntosh, L. P.; Hand, G.; Johnson, P. E.; Joshi, M. D.; Korner, M.; Plesniak, L. A.; Ziser, L.; Wakarchuk, W. W.; Withers, S. G. Biochemistry 1996, 35, 9958).
27. Kempton, J. B.; Withers, S. G. Biochemistry 1992, 31, 9961.
28. Prade, H.; Mackenzie, L. F.; Withers, S. G. Carbohydr. Res. 1998. In press.
29. α-Glycosyl fluorides are synthesised by reaction of the per-O-acetylated sugar with HF/pyridine followed by deprotection with methoxide in methanol.
30. α-Glucosyl fluoride does not function as a substrate for, or even bind to, the wild-type enzyme. The axial fluorine should also disfavor binding to the aglycon site, decreasing competition between the donor and acceptor sugars.
31. Day, A. G.; Withers, S. G. Can. J. Biochem. 1986, 64, 914.
32. Such β(1,3) transfer to xylose has been seen with other glycosidases (Lopez, R.; Fernandez-Mayoralas A. J. Org. Chem. 1994, 59, 737.).
33. The yield of the disaccharide was maximized by adding only 0.5-0.75 equiv of the glycosyl donor.